【摘要】：背景:研究證實(shí),維生素C除了具有抗氧化作用外,還具有促進(jìn)細(xì)胞增殖的重要作用,但至今為止,有關(guān)維生素C對高糖環(huán)境下人脂肪干細(xì)胞的增殖作用及相關(guān)分子機(jī)制尚不明確。目的:研究維生素C促進(jìn)高糖環(huán)境下人脂肪干細(xì)胞增殖的作用及相關(guān)分子機(jī)制。方法:取高塘環(huán)境中培養(yǎng)的第3代人脂肪干細(xì)胞,分別以0,100,150,200,250,300μmol/L維生素C處理,以低糖環(huán)境中培養(yǎng)的細(xì)胞為對照,MTT法篩選最佳維生素C干預(yù)濃度及處理時(shí)間,進(jìn)行Western Blot檢測。EdU檢測:將高糖環(huán)境中培養(yǎng)的人脂肪干細(xì)胞分4組培養(yǎng),空白對照組不進(jìn)行任何干預(yù),維生素C組加入最佳濃度維生素C,LY294002組加入PI3K/AKT信號通路抑制劑LY294002與最佳濃度維生素C,U0126組加入ERK/MAPK信號通路抑制劑U0126與最佳濃度維生素C,以低糖環(huán)境中培養(yǎng)的細(xì)胞為對照,干預(yù)48 h后,檢測Ed U標(biāo)記陽性細(xì)胞數(shù)。結(jié)果與結(jié)論:(1)CCK-8法檢測:200μmol/L維生素C促進(jìn)高糖環(huán)境下人脂肪干細(xì)胞的增殖作用最強(qiáng),所以此為最佳干預(yù)濃度,最佳處理時(shí)間為48 h;(2)Western Blot檢測:對照組、0μmol/L維生素C處理組、200μmol/L維生素C處理組p-AKT蛋白表達(dá)無差異,200μmol/L維生素C處理組p-ERK蛋白表達(dá)高于0μmol/L維生素C處理組(P0.01);(3)Ed U檢測:維生素C組、LY294002組、對照組EdU標(biāo)記陽性細(xì)胞數(shù)高于空白對照組(P0.01),維生素C組EdU標(biāo)記陽性細(xì)胞數(shù)高于U0126組(P0.01);(4)結(jié)果表明:維生素C通過ERK/MAPK信號通路促進(jìn)高糖環(huán)境下人脂肪干細(xì)胞的增殖。
[Abstract]:Background: studies have confirmed that vitamin C not only has antioxidant effects, but also plays an important role in promoting cell proliferation. The effect of vitamin C on the proliferation of human adipose stem cells in high glucose environment and its molecular mechanism are not clear. Aim: to study the effect of vitamin C on proliferation of human adipose stem cells in high glucose environment and its molecular mechanism. Methods: the third generation of human adipose stem cells cultured in Gaotang was treated with 0100150200250300 渭 mol/L vitamin C, and the cells cultured in low-sugar environment were used as control. The best intervention concentration and treatment time of vitamin C were screened by MTT method. Western Blot detection. EdU detection: human adipose stem cells cultured in high sugar environment were divided into 4 groups, the blank control group did not intervene, vitamin C group added the best concentration of vitamin C, LY294002 group added PI3K/AKT signaling pathway inhibitor LY294002 and best concentration of vitamin C, U0126 group added ERK/MAPK signal pathway inhibitor U0126 and optimum concentration of vitamin C, and cultured cells in low glucose environment were treated for 48 h. The number of Ed U labeled positive cells was detected. Results and conclusion: (1) CCK-8 assay showed that 200 渭 mol/L vitamin C had the strongest effect on the proliferation of human adipose stem cells in high glucose environment, so this was the best intervention concentration and the best treatment time was 48 hours. (2) Western Blot: there was no difference in p-AKT protein expression among control group, 0 渭 mol/L vitamin C treatment group and 200 渭 mol/L vitamin C treatment group. The expression of p-ERK protein in 200 渭 mol/L vitamin C treatment group was higher than that in 0 渭 mol/L vitamin C treatment group (P0.01). (3) Ed U detection: the number of EdU labeled positive cells in vitamin C group, LY294002 group and control group was higher than that in blank control group (P0.01), and the number of EdU labeled positive cells in vitamin C group was higher than that in U0126 group (P0.01). (4) the results showed that vitamin C promoted the proliferation of human adipose stem cells in high glucose environment through ERK/MAPK signaling pathway.
【作者單位】： 廣東醫(yī)科大學(xué)附屬醫(yī)院整形外科研究所;
【基金】：2015年國家自然科學(xué)基金面上項(xiàng)目(81570260) 2014年廣東省自然科學(xué)基金(2014A030313535) 2016廣東省科技計(jì)劃項(xiàng)目(2016A020214018) 2014湛江市財(cái)政資金科技專項(xiàng)經(jīng)費(fèi)資助項(xiàng)目(2014C01022) 2015廣東醫(yī)科大學(xué)博士基金(BJ201510) 2015年廣東醫(yī)科大學(xué)附屬醫(yī)院博士啟動基金(BJ201505)~~
【分類號】：R587.2

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